EP0987272A1 - Verfahren zur Herstellung von Polyguluronicsäuren - Google Patents

Verfahren zur Herstellung von Polyguluronicsäuren Download PDF

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Publication number
EP0987272A1
EP0987272A1 EP99114083A EP99114083A EP0987272A1 EP 0987272 A1 EP0987272 A1 EP 0987272A1 EP 99114083 A EP99114083 A EP 99114083A EP 99114083 A EP99114083 A EP 99114083A EP 0987272 A1 EP0987272 A1 EP 0987272A1
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acid
solution
polyguluronic
alginic acid
acids
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French (fr)
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EP0987272B1 (de
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William Alan Marritt
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Seiko Epson Corp
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Seiko Epson Corp
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H7/00—Compounds containing non-saccharide radicals linked to saccharide radicals by a carbon-to-carbon bond
    • C07H7/02—Acyclic radicals
    • C07H7/033—Uronic acids
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0084—Guluromannuronans, e.g. alginic acid, i.e. D-mannuronic acid and D-guluronic acid units linked with alternating alpha- and beta-1,4-glycosidic bonds; Derivatives thereof, e.g. alginates

Definitions

  • the present invention relates to a process for the manufacture of polyguluronic acids having degrees of polymerization less than 20 and substantially free of mannuronic acid contamination.
  • Polyguluronic acids because of their high affinity for calcium ions, are expected to have utility as scale inhibitors and scale deposit removers. The biodegradability of polyguluronic acids makes them particularly valuable with respect to environmental acceptability and waste disposal. Additionally, polyguluronic acid derivatives, in which a hydrophobic polymer is covalently attached to the reducing terminus of the polyguluronic acid, are useful as dispersants in pigment dispersed aqueous ink compositions for use in ink jet printing. Furthermore, polyguluronic acids, having low degrees of polymerization, have been shown to exhibit root-growth promoting activity in barley (M.
  • Alginic acids from which polyguluronic acids can be obtained, are unbranched polymers of 1 ⁇ 4 linked ⁇ -L-guluronic acid (G) and ⁇ -D-mannuronic acid (M) of varying proportions and sequence.
  • G 1 ⁇ 4 linked ⁇ -L-guluronic acid
  • M ⁇ -D-mannuronic acid
  • a typical structure of an alginic acid molecule may be represented schematically as follows:
  • Alginic acids are best described as block copolymers in which there are polyguluronic acid sequences (G-blocks), polymannuronic acid sequences (M-blocks), and sequences containing random arrangements of both guluronic acid and mannuronic acid (MG-blocks).
  • alginates which are salts of alginic acid
  • hydrolysis products can be separated to give two predominantly homopolymeric fractions, polyguluronic acid and polymannuronic acid.
  • the most often cited procedure for the preparation of the sodium salt of polyguluronic acid is a heterogeneous acidic hydrolysis method disclosed in A. Haug et al., "Studies on the Sequence of Uronic Acid Residues in Alginic Acid," Acta Chemica Scandinavica, 21, 691-704 (1967).
  • the acidic hydrolysis described therein requires that one part of sodium alginate be suspended in twenty parts of 0.3 M hydrochloric acid solution.
  • the hydrolysis is a heterogeneous reaction.
  • the heterogeneous mixture is heated for 10+ hours at 100°C and then the solid is separated from the acidic solution by centrifugation or filtration. After the collected solid is dissolved in water by neutralizing with dilute sodium hydroxide solution, twenty parts of 0.3 M hydrochloric acid solution are added to the solution resulting in reprecipitation of the partially hydrolyzed alginic acid.
  • the resulting heterogeneous mixture is heated for an additional 10+ hours at 100°C and the solid product is again separated from the acidic solution by centrifugation or filtration.
  • the collected solid is dissolved in water by neutralizing with dilute sodium hydroxide solution and then sodium chloride and water are added to yield a solution which is 0.5 wt% alginic acid and O.1 M sodium chloride.
  • An approximately equal volume of 0.025 M hydrochloric acid solution is added to the alginic acid salt solution until a pH value of 2.85 is obtained.
  • the precipitated solid is separated from the acidic solution by centrifugation or filtration.
  • the isolated solid is dissolved in water by neutralizing with dilute sodium hydroxide solution and then precipitated with excess ethanol.
  • the precipitated solid is washed with ethanol, washed with ether, and dried.
  • the sodium salt of the polyguluronic acid prepared by this heterogeneous acidic hydrolysis method has an average degree of polymerization between 15 and 20.
  • the mannuronic acid content is between 5 and 15%, and the yield of product is between 15 and 20%.
  • heterogeneous acidic hydrolysis method of A. Haug et al. is useful for laboratory scale preparations of polyguluronic acid, it would be difficult to implement on a larger scale, such as that which would be used in industrial production. This is because in the separation step of that method, the concentration of alginic acid is only a very dilute 0.25 wt.%. Additionally, the method has multiple steps and is complicated.
  • the homogeneous acidic hydrolysis method of A. Haug et al. may be useful for laboratory scale preparations of polyguluronic acid, it also would be difficult to implement on a larger scale.
  • the concentration of sodium alginate in the hydrolysis step is only a very dilute 0.5 wt.%. Although one can easily conceive of increasing the concentration of sodium alginate, in practice this cannot be done. At concentrations slightly greater than 0.5 wt.%, sodium alginate does not remain soluble throughout the hydrolysis reaction in the range of pH values at which the acidic hydrolysis is effected.
  • the species which precipitate in the course of the hydrolysis are incompletely hydrolyzed and, although rich in guluronic acid, they contain significant amounts of mannuronic acid.
  • the incompletely hydrolyzed species have degrees of polymerization greater than 20. Because the homogeneous acidic hydrolysis conditions are milder than those described above in the heterogeneous acidic hydrolysis procedure, the incompletely hydrolyzed species are not further hydrolyzed after precipitating from solution.
  • alginate salts of organic bases are quite soluble in aqueous solution. Furthermore, the present inventor has found that these salts remain soluble in aqueous solution throughout a hydrolysis reaction in the range of pH values at which the hydrolysis is effected at a reasonably fast rate. The present invention has been made based on such findings.
  • a process for the manufacture of polyguluronic acids having degrees of polymerization less than 20 and substantially free of mannuronic acid contamination, comprising the steps of:
  • Polyguluronic acids obtained by the process of the present invention have degrees of polymerization less than 20 and are substantially free of mannuronic acid contamination.
  • substantially free of mannuronic acid contamination means that the mannuronic acid content is less than about 8 wt.%, and preferably less than about 5 wt.%.
  • homogeneous acidic hydrolysis in which sodium alginate is used as the starting material, cannot be carried out at concentrations significantly greater than 0.5 wt.%.
  • alkali metal salts of alginic acid including lithium alginate, potassium alginate, rubidium alginate, and cesium alginate, and ammonium alginate.
  • Alginate salts of metals other than alkali metals are insoluble or sparingly soluble in aqueous solution and, thus, cannot be used in a homogeneous acidic hydrolysis.
  • alginate salts of organic bases are very advantageous in that they are quite soluble in aqueous solution and remain soluble in aqueous solution throughout a hydrolysis reaction in the range of pH values at which the hydrolysis is effected at a reasonably fast rate.
  • organic bases usable in the present invention include organic amines (for example, mono-, di- and trialkyl (preferably C 1-8 alkyl, more preferably C 1-6 alkyl) amines, tetraalkyl (preferably C 1-8 alkyl, more preferably C 1-6 alkyl) ammonium hydroxides, and aromatic amines), tetraarylphosphonium hydroxide, tetraarylarsonium hydroxide, and tetraarylstibonium hydroxide.
  • organic amines for example, mono-, di- and trialkyl (preferably C 1-8 alkyl, more preferably C 1-6 alkyl) amines, tetraalkyl (preferably C 1-8 alkyl, more preferably C 1-6 alkyl) ammonium hydroxides, and aromatic amines), tetraarylphosphonium hydroxide, tetraarylarsonium hydroxide, and tetraarylstibonium hydroxide.
  • the organic base has a relatively low molecular weight for two reasons.
  • low molecular weight organic bases are appreciably soluble in water even before protonation.
  • low molecular weight organic bases yield salts of alginic acid for which the viscosity of an aqueous solution is less than that of a comparable solution prepared using a higher molecular weight organic base.
  • organic bases examples include methylamine, dimethylamine, trimethylamine, tetramethylammonium hydroxide, ethylamine, diethylamine, triethylamine, tetraethylammonium hydroxide, n-propylamine, di-n-propylamine, tri-n-propylamine, tetra-n-propylammonium hydroxide, isopropylamine, di-isopropylamine, N-methyl-ethylamine, N-methyl-n-propylamine, N-methyl-isopropylamine, N,N-dimethyl-ethylamine, N, N-dimethyl-n-propylamine, N, N-dimethyl-isopropylamine, N, N-diethyl-n-propylamine, N, N-diethyl-isopropylamine, N, N-di-n-propyl-isopropylamine, N, N-
  • the family of methylamines - methylamine, dimethylamine, trimethylamine, and tetramethylammonium hydroxide - are used because the viscosities of the resulting solutions of alginic acid salts are relatively low.
  • the alginic acid to be used as a starting material in the present invention may be any alginic acid. Further, any commercially available alginic acid may be used. Because polyguluronic acid is the target product, an alginic acid, which is rich in guluronic acid, is preferred. For example, alginic acids extracted from the seaweeds Laminaria hyperborea and Lessonia flavicans are particularly rich in guluronic acid, having guluronic acid to mannuronic acid ratios of approximately two to one.
  • an alginic acid having a low average molecular weight is preferred as a starting material.
  • an alginic acid having a molecular weight of less than or equal to 50,000 g/mole is used.
  • Sodium salts of alginic acid having average molecular weights of 10,000 g/mole are at present commercially available from Kimitsu Chemical Industries (Product Name: ULV-LlG) and Fuji Chemical Industries (Product Name: Snow Algin ULV-Sl).
  • a particularly useful alginic acid starting material can be readily obtained by acidifying to a pH value of 2 or less a solution of either of these sodium salts and then collecting the solid by filtration or centrifugation.
  • the alginic acid is dissolved in water by neutralization with an organic base to yield a solution containing 5 wt.% or more of alginic acid.
  • an organic base may be used to effect the neutralization with the pH of the resulting solution being in the range of about 5 to about 9.
  • the resulting solution will generally contain small amounts of insoluble impurities which can be easily removed by filtration prior to the acidic hydrolysis step.
  • the alginic acid is hydrolyzed to low molecular weight components including polyguluronic acids.
  • the hydrolysis is carried out under acidic conditions, preferably with heating.
  • the pH value is preferably in the range of 3.2 to 5.0, more preferably in the range of 3.5 to 4.2.
  • This acidity may be realized by the addition of any water-soluble acid, and examples of preferred acids usable herein include hydrochloric acid and lactic acid.
  • the pH value of the solution is continuously adjusted during the hydrolysis. This is because it has been found that the pH of the solution increases gradually during the course of the hydrolysis.
  • Adjustment of the pH can be carried out through the use of commercially available metering devices. According to a preferred embodiment of the present invention, such devices use a pH electrode as a sensor to provide feedback to one or more syringe pumps, which add acid or base to maintain the pH of the solution at a set value.
  • a temperature of 80°C or above is preferred for carrying out the hydrolysis reaction. Although temperatures less than 80°C may be used, the rate of hydrolysis will be correspondingly slower.
  • the hydrolysis reaction may also be carried out in pressure vessels, such that temperatures greater than the boiling point of water may be used.
  • a temperature of less than about 120°C is preferred because nonselective decomposition of the polysaccharide products, in competition with the hydrolysis reaction, is significant at temperatures greater than about 120°C.
  • the hydrolysis step may be carried out under an ambient atmosphere or an inert atmosphere, with an inert atmosphere being preferred because air oxidation of the products will be minimized.
  • An inert atmosphere of high purity nitrogen is preferred from the standpoint of cost.
  • the reaction is allowed to proceed for a time period sufficient to complete the hydrolysis.
  • the time period which is sufficient to hydrolyze the alginic acid to lower molecular weight components including polyguluronic acids depends on several factors.
  • One factor is the temperature of the reaction.
  • a second factor is the initial concentration of the alginic acid.
  • a third factor is the average molecular weight of the starting material alginic acid.
  • a fourth factor is the pH value of the hydrolysis solution.
  • a variety of analytical techniques may be used. Most simply, the viscosity of the hydrolysis solution may be monitored and the reaction judged to be complete when a stable value is obtained.
  • aliquots of solution may be removed periodically, the polyguluronic acid fraction isolated, and the polyguluronic acid fraction analyzed by a variety of techniques.
  • Useful techniques included 1 H-NMR, gel permeation chromatography, and low angle light scattering.
  • the pH range of the acidification step in which polyguluronic acids are selectively precipitated, specifically refers to a pH range in which polyguluronic acids are selectively precipitated from polymannuronic acids.
  • the pH value is preferably in the range of 3.0 to 3.6, more preferably in the range of 3.1 to 3.5.
  • the reported pKa values of polyguluronic acid and polymannuronic acid are 3.65 and 3.38, respectively. Based on this difference, acidification in the pH range of the present invention, results in selective precipitation of polyguluronic acids from polymannuronic acids, which remain in solution.
  • the carboxylic acid groups of polyguluronic acid may not be protonated and some organic base salts may coprecipitate with the polyguluronic acid. If this is problematic for the application in which the polyguluronic acid is to be used, the selectively precipitated polyguluronic acid may be isolated, dissolved in water and then reprecipitated by acidification to a pH value less than about 2.
  • the acidification for the precipitation of polyguluronic acid may be realized by the addition of any water-soluble acid. Examples of preferred acids include hydrochloric acid and lactic acid.
  • the polyguluronic acids are separated from the acidified solution by conventional methods. Such methods include filtration and centrifugation.
  • the product may then be washed with acidified water and/or an aqueous alcohol solution and then dried by conventional methods.
  • the polyguluronic acids prepared according to the method of the present invention have degrees of polymerization of less than 20 and contain less than 8% mannuronic acid as determined by 1 H-NMR.
  • a sample for NMR analysis is prepared by dissolving the polyguluronic acid in deuterium oxide (D 2 0) by neutralization with a solution of sodium deuteroxide (NaOD) in D 2 0.
  • NaOD sodium deuteroxide
  • the H-l (internal) peaks of guluronic acid and mannuronic acid in polyuronic acids occur at 5.05 ppm and 4.67-4.70 ppm, respectively, relative to the internal reference standard, sodium 3-(trimethylsilyl)propionate-d 4 .
  • the amount of mannuronic acid impurity in the polyguluronic acid product can be calculated.
  • the H-l (reducing end) peaks of the ⁇ and ⁇ anomers of polyguluronic acid and polymannuronic acid occur at 5.21 ppm and 4.84-4.89 ppm, respectively, relative to the internal reference standard, sodium 3-(trimethylsilyl)propionate-d 4 .
  • the degree of polymerization of the polyguluronic acid product can be calculated.
  • Alginic acid used in the following specific example was prepared from a commercial sodium alginate (Kimitsu Chemical Industries; Product Name: ULV-LlG; average molecular weight: 10,000). 200 g of ULV-LlG was dissolved in 1.8 liters of deionized water with stirring. While stirring the solution, concentrated hydrochloric acid was added dropwise which resulted in precipitation of alginic acid. The addition of hydrochloric acid was continued until the pH of the mixture reached a value of 0.90. The mixture was stirred for an additional 4 hours and then set aside to stand for 12 hours. Most of the clear supernatant was discarded and the remaining mixture was filtered through a #2 Whatman filter paper to collect the off-white solid. The solid was air-dried for several days and then further dried under vacuum to a constant weight.
  • the solution was heated to 90°C with stirring and then 1 M hydrochloric acid solution was added dropwise to the solution until the pH reached a value of about 3.9, as determined using Hydrion Microfine pH test paper having a range of 2.9 to 5.2.
  • a reflux condenser to which a nitrogen inlet line and a bubbler were attached, was inserted into one of the necks of the flask and a glass stopper was inserted into the other neck of the flask.
  • the solution was then heated at reflux for 12 hours. During this time period, the pH was periodically sampled through the inlet which was closed with the glass stopper and 1 M hydrochloric acid solution was added as needed to maintain the pH of the solution at a value of about 3.9.
  • the pH was periodically sampled through the inlet which was closed with the glass stopper and 1 M hydrochloric acid solution was added as needed to maintain the pH of the solution at a value of about 3.7.
  • the solution was allowed to cool to room temperature and then acidified to a pH value of 3.3 using 1 M hydrochloric acid solution.
  • the mixture was set aside for 12 hours during which time the solid polyguluronic acid settled to the bottom of the flask.
  • the pH value was measured again and additional 1 M hydrochloric acid was added in order to readjust the pH value to 3.3.
  • the solid was collected by vacuum filtration on a fine porosity fritted glass filter and then washed several times with 50 wt.% aqueous methanol solution.
  • the 1 H-NMR spectrum was measured at 80°C and the peaks corresponding to the H-l (internal) peak of guluronic acid in polyuronic acids, the H-l (internal) peak of mannuronic acid in polyuronic acids, the H-l (reducing end) peak of the ⁇ anomers, and the H-l (reducing end) peak of the ⁇ anomers were integrated.
  • the integration values are given in Table 1 as shown below. This region of the spectrum is shown in Figure 1. Proton Chemical Shift (ppm) (vs.

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EP99114083A 1998-07-21 1999-07-20 Verfahren zur Herstellung von Polyguluronsäuren Expired - Lifetime EP0987272B1 (de)

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JP20502598A JP3932679B2 (ja) 1998-07-21 1998-07-21 ポリグルロン酸の製造法
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Cited By (1)

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Publication number Priority date Publication date Assignee Title
WO2003099870A3 (fr) * 2002-05-29 2004-04-01 Ct D Etude Et De Valorisation Procedes d'obtention d'oligo-mannuronates et guluronates, les produits obtenus et leurs utilisations

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AU2003252735A1 (en) * 2002-07-30 2004-02-16 Seiko Epson Corporation Polyuronic acid derivative and aqueous ink composition comprising polyuronic acid derivative
GB2430881B (en) 2005-10-06 2010-10-13 Ntnu Technology Transfer As Oligoelectrolyte polyols for the treatment of mucosal hyperviscosity
GB0707096D0 (en) 2007-04-12 2007-05-23 Ntnu Technology Transfer As Method
GB0904941D0 (en) 2009-03-23 2009-05-06 Ntnu Technology Transfer As Composition
GB0904942D0 (en) 2009-03-23 2009-05-06 Ntnu Technology Transfer As Composition
JP2011241154A (ja) * 2010-05-14 2011-12-01 Tokuyama Dental Corp アルギン酸類含有水性組成物
JP2012121963A (ja) * 2010-12-07 2012-06-28 Meiji Univ ポリグルロン酸の製造方法、ポリマンヌロン酸の製造方法、ポリグルロン酸、及びポリマンヌロン酸
JP6128545B2 (ja) * 2012-11-13 2017-05-17 花王株式会社 容器
JP2016111957A (ja) * 2014-12-12 2016-06-23 伊那食品工業株式会社 多糖類水溶液の製造方法、それによって得られた多糖類水溶液及び多糖類水溶液を含む食品

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US4689322A (en) * 1982-07-28 1987-08-25 Algina Aktiengesellschaft Pharmaceutical products, calcium mixed salts of polymeric, anionic carboxylic acids and/or their esters of sulfuric acid, and methods for their preparation and use
JPH06217774A (ja) * 1992-12-04 1994-08-09 Kibun Food Chemifa Co Ltd 新規アルギン酸分解酵素、その製造方法およびこれを用いたポリマンヌロン酸の製造方法

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003099870A3 (fr) * 2002-05-29 2004-04-01 Ct D Etude Et De Valorisation Procedes d'obtention d'oligo-mannuronates et guluronates, les produits obtenus et leurs utilisations

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ATE238324T1 (de) 2003-05-15
US6770756B1 (en) 2004-08-03
DE69907104D1 (de) 2003-05-28
DE69907104T2 (de) 2003-10-30
EP0987272B1 (de) 2003-04-23
JP3932679B2 (ja) 2007-06-20

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